ES2198580T3 - NEW SALTS OF ANIONIC COMPOUNDS OF RU (III), USED AS ANTIMETASTASIC AND ANTINEOPLASTIC AGENTS. - Google Patents

Abstract

THE INVENTION REFERS TO AN ANIONIC COMPLEX OF RU (III) WITH AN AMONIUM CATION OF FORMULA (I), WHERE R 1, R 2 YR 3, EQUAL OR DIFFERENT BETWEEN, ARE SELECTED FROM THE GROUP FORMED BY H, RENT C 1 - C 6 LINEAR OR RAMIFIED, SATURATED OR UNSATURATED, CYCLEALKYL C 3 C 7, PHENYL AND ARYL; OR NR 1 R 2 R 3 IS A HE TEROCICLO CONTAINING NITROGEN, OF 5-7 MEMBERS, SATURATED OR UNSATURATED, OPTIONALLY CONTAINING ONE OR MORE ATOMS OF O, SY / ON, BEING OPTIONALLY REPLACED SUCH NITROGEN ATOMO BY A RESIDUE OF C 1 - C 4 RENT, ARILO OR BE NCILO; SUCH HETEROCICLE CONTAINING NITROGEN, MAY BE OPTIONALLY CONDENSED WITH A BENZO GROUP AND / OR REPLACED BY C 1 - C 4, ALCOXYL C 1 - C 4, C 1 - C 4, ARILO OR BENCILO; WHERE R SU B, 4 YR 5, EQUAL OR DIFFERENT BETWEEN, ARE SELECTED FROM THE GROUP FORMED BY H, RENT C 1 - C 6, CYCLEALKYL C 3 - C 7, PHENYL AND ARYL, OR 4 YR 5 FORMAN, TOGETHER WITH THE ATOM OF S, A HETEROCICLE OF 4-7 MEMBERS. IN ADDITION, THE PROCEDURE FOR THE PREPARATION OF SALES ACIDS, THE COMPOSITIONS CONTAINING THEM, AND ITS USE AS ANTIMETASTATIC AND ANTHINEOPLASTIC AGENTS, IS DESCRIBED.

Description

New salts of anionic compounds of Ru (III), used as antimetastatic and antineoplastic agents.

Field of the Invention

The present invention relates to new salts of anionic compounds of Ru (III) with ammonium cations that are particularly useful as antimetastatic agents and antineoplastic

State of the art

The discovery of the properties cisplatin antineoplastic (cis-diaminodichloroplatin (II)), used frequently today for both monochemotherapy as for antitumor polychemotherapy, it has aroused interest by the study of the antitumor activity of the compounds metalorganic Like other antitumor agents, the specificity of action of cisplatin is directed at those neoplasms that affect specific compartments (testicles, ovary, bladder, head and neck), while other types of tumors such as those located in the lungs and chest and the colorectal tumor are little sensitive to this treatment Pharmacist (C.F.J. Barnard et al. Chemistry in Britain, 1001-1004, 1986).

Thus, research in the field of coordinated compounds intends to develop new medications that contain Pt and / or other transition elements that allow increased use spectrum, as well as obtaining levels of toxicity lower than those generated by antitumor agents known thanks to the state of the art.

The potentiality of the use of ruthenium as an agent antitumor against the platinum alternative has been studied in compounds of Ru (II), such as the cis-tetrakis-dimethylsulfoxide Ruthenium (II) (T. Giraldi et al., Cancer Res., 37, 2662, 1977) and, more recently, in neutral Ru (III) compounds such as fac- [RuCl 3 (NH 3) 3] (M.J. Clarke, Metal lons in Biological Systems, 11 (5), 231-281, 1980, Helmut Sigel Ed.).

In addition, several compounds have been prepared Ru (III) anionics with five-member heterocycles, in particular the imidazolium-bis-imidazol-tretrachloro-ruthenate (III) (B.K. Keppler et al., J. Cancer Res. Clin. Oncol, 111: 166-168, 1986).

Recently, some Ru (III) compounds have been developed with DMSO; in particular, international patent application WO 90/13553 describes some Ru (III) compounds represented by formula (i): [Ru (R_ {x} R_ {Y} SO) \; (Cl_ {3}) AB] \ eqnum {(i)} where R x R y SO is a sulfoxide, preferably dimethylsulfoxide; A is a sulfoxide or a chloride and B is a nitrogen ligand selected from the group consisting of ammonia, primary, secondary and tertiary amines and heterocycles containing nitrogen atoms.

These compounds, which can be used as agents antineoplastic, they are neutral when A is a sulfoxide, while they have a negative charge when A is a chloride. In this last case, the anionic compounds are isolated in the form of salts corresponding with alkaline and alkaline earth cations, preferably sodium.

Despite their antimetastatic activity, the compounds mentioned above show some drawbacks serious that make administration and formulation in therapeutically effective compositions. Fundamentally, these anionic compounds, when isolated in Sodium salts form, always contain two solvent molecules of crystallization and cannot be isolated in pure form, that is, Without these crystallization molecules.

In particular, the [trans-RuCl_ {4} (DMSO) Im] Na 2DMSO, which corresponds to the formula (i) where Rx = Ry = methyl, A = Cl, B = imidazole and whose negative charge is neutralized with Na +, is able to exercise a good antineoplastic activity and antimetastatic on tumor models in mice. It is also Salt is characterized by its high water solubility, which facilitates Your administration

However, this compound can only be isolated with two DMSO dissolution molecules, as shown in the examples described in the international patent application mentioned above (in particular, in examples 3, 4, 8 and 10), which reduces the stability of the compound.

In addition, a variability can be observed qualitative of the crystallization molecules, which can be DMSO, acetone and water and, more frequently, two molecules of dimethylsulfoxide (DMSO) (E. Alessio et al. Inorganic Chimica Acta, 203 (1993): 205-217).

The variation of the crystallization molecules makes it difficult to control from one preparation to another, which raises unknowns in structural analysis, in weight determination molecular and in elemental analysis. Therefore, they can appear problems related to the purity of the compound and its subsequent formulation. Precisely, the poor reproducibility structural of these compounds implies that the same amount of Ruthenium does not always correspond to the dose administered. Thus, the non-constant dose treatment and therapeutic activities that cannot be easily predicted reduce the pharmaceutical interest of these compounds.

Another drawback presented by the compound mentioned above derives from the negative effects Pharmacists caused by DMSO, introduced into the body in double or equimolar amounts with respect to the compound of Ruthenium, and its differentiation properties, as Klaas reports Kramer et al. (Gen. Pharmac., Vol. 26, n2 6, pp. 1403-1497, 1995).

Finally, as the salts mentioned previously contain dissolution molecules, they degrade quickly when exposed outdoors or at temperature ambient to provide a semi-solid brown substance.

Summary

The applicant has discovered, so unusual, new salts of anionic compounds of Ru (III) with ammonium cations that show great antineoplastic activity and antimetastatic These salts are represented by the formula (I):

one

where R 1, R 2 and R 3, the same or different from each other, they are selected from a group formed by H, C 1 -C 6 alkyl, linear or branched, saturated or unsaturated, cycloalkyl C 3 -C 7, phenyl and aryl;

or NR 1 R 2 R 3 is a heterocycle of 5-7 members nitrogen, saturated or unsaturated, containing, optionally, one or more atoms of O, S and / or N; being said nitrogen atom optionally substituted by a residue C 1 -C 4 alkyl, an aryl or a benzyl; said nitrogen-containing heterocycle optionally condensing  with a benzyl group and / or substituted by an alkyl group C 1 -C 4, alkoxy C 1 -C 4, aryl or benzyl;

where R_ {4} and R_ {5}, same or different each other, they are selected from the group formed by an atom of H, C 1 -C 6 alkyl, cycloalkyl C 3 -C 7, phenyl and aryl or R 4 and R 6 form, together with the atom of S, a heterocycle of 4-7 members.

Another object of the present invention is provide a new procedure for the preparation of previous salts (I) comprising the synthesis of [trans-RuCl 4 (R 4 R 5 SO) 2] [(R 4 R 5 SO) H] (II), where R 4 and R 5 have the meaning indicated above, which is obtained by treatment of RuCl 3 with R 4 -SO-R 5 in the presence of HCl, and the subsequent reaction of compound (II) with a compound nitrogenous of formula NR 1 R 2 R 3, in the presence of a organic solvent

In addition, the present invention relates to pharmaceutical compositions containing a concentration Therapeutically effective of at least one of the above salts of formula (I), in combination with excipients and / or diluents suitable and its use as antimetastatic agents and / or antineoplastic

Description of the figures

Figure A1 and Figure A2 show, respectively, the external connective capsule and an area of tumor tissue necrosis of control mice affected by breast carcinoma (MCa).

Figures B1 and B2 show, respectively, the external connective capsule and a zone of tissue necrosis tumor of mice affected by breast carcinoma (MCa), after treatment with [trans-RuCl_ {4} (DMSO) Im] Na 2DMSO, as described in the international patent application WO 90/13553, mentioned above.

Finally, Figures Cl and C2 show the external connective capsule and a zone of tissue necrosis tumor of mice affected by breast carcinoma (MCa), after salt treatment [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example 2) of the present invention.

Detailed description of the invention

The characteristics and advantages of this invention will be better illustrated throughout the following detailed description.

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In the salts of formula (I), mentioned above, the nitrogen atom of the nitrogen compound of formula NR 1 R 2 R 3 contributes with its long torque to the formation of the coordination bond with the atom of Ru (III).

More specifically, in the salts of the invention, R1 is an atom of H or an ethyl and is, in turn, same as R2 and R3.

When NR 1 R 2 R 3 is a heterocycle of five members containing nitrogen, is selected from the group formed by imidazole, N-methyl-imidazole, pyrazole and oxazole; preferably, this nitrogen heterocycle is the imidazole

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3,5-lutidina y 4-metilpiridina.When NR 1 R 2 R 3 is a 6-membered heterocycle, it is selected from the group consisting of pyridine, pyrazine,

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3,5-lutidine and 4-methylpyridine.

When NR 1 R 2 R 3 is a heterocycle of 7 members, it is preferably selected from the group consisting of azepine, diazepine and oxazepine.

Finally, when said heterocycle is condensed with a benzyl group, it is preferably selected from the group formed by indazole, isoquinoline, benzimidazole and 1,5,6-trimethyl-benzimidazole.

In the salts of the invention of formula (I), the ligand sulfoxide, R 4 -SO-R 5, is, preferably, dimethylsulfoxide (R 4 = R 5 = methyl), diethylsulfoxide (R 4 = R 5 = ethyl) or tetramethylene sulfoxide (together with the atom of S, R 4 and R 5 form a ring of five members).

These salts may unexpectedly exert a antitumor activity and, in particular, antimetastatic, significantly higher than the corresponding salts of sodium, when applied in equimolar doses and models of equitable treatment

Also, the salts of formula (I), with respect to those described in the state of the art are much more favorable for its high stability in contact with air, while the corresponding sodium salts, extremely hygroscopic, have a great tendency to hydrolysis. In addition, the salts of the invention show no pharmaceutical drawbacks with regarding the known complexes, due to the presence of DMSO crystallization molecules.

Although the salts of the formula (I) do not have crystallization molecules, have constant wave values average, as well as reproducible analytical results. This allows that the formulations contain defined amounts of the compound active and treatments with constant doses by nature.

The salts of the Ru (III) compounds of formula (I) are obtained by a new procedure particularly simple and favorable comprising the following stages:

one)

reactions of RuCL_3 with R4 -SO-R5 in presence of HCl, to produce the compound of formula (II)

two

where R_ {4} and R_ {5} have the meaning before indicated.

At this stage of the reaction, the substituents R 4 and R 5 are preferably methyl, that is, the RuCl 3 reacts with dimethylsulfoxide (DMSO) to produce [(Me 2 SO) 2 H] [trans-RuCl 4 (Me 2 SO) 2].

According to the form of preparation chosen, RuCl 3 is previously dissolved in an organic solvent heated, preferably ethanol or methanol; the R4 -SO-R5 and HCl concentrate are added to the solution obtained by this procedure and heated to a temperature that preferably varies between 60 ° C and 90 ° C and even more preferably at a temperature approximately 80ºC.

two)

The compound (II), obtained from step (1), reacts at room temperature with a nitrogenous compound of formula NR 1 R 2 R 3, in molar ratios ranging from 1: 2 to 1: 6, in one or more organic solvents, preferably acetone or dichloromethane, for the production of the salts of formula (I), according to the scheme described then:

3

where R_ {1} -R_ {5} have the meaning before indicated.

In step (2), the intermediate product (II) reacts with the nitrogen compound of formula NR, R 2 R 3, causing the replacement of one of the two axial sulfoxide groups, the protonization of a molecule of nitrogen compound and the final release of two molecules of sulfoxide crystallization. Thus, the compounds (I) are obtained With very high yields.

Another object of the present invention is the use of the above salts (I) for the treatment of neoplasms of different nature and for the prevention of the formation of metastasis. These neoplasms are preferably solid tumors, such as carcinoma of the gastroinensayoinal tract, carcinoma of breast, lung tumors, metastatic carcinoma and pulmonary metastases of metastatic tumors.

The salts of the invention can be administered, favorably, parenterally, orally, topically or transdermal

Among the forms of administration via parenteral, intravenous, intramuscular route is preferable, intraperitoneal and subcutaneous.

Treatment with these salts varies depending on of the route and method of administration, as well as the severity of the neoplasms; in addition, it varies in relation to age, the weight of the body and the patient's general state of health.

The therapeutically effective dose of these salts, to be administered in single or multiple doses, varies preferably between 0.1 and 300 mg / kg / day and even more preferably, between 10-200 mg / kg / day when you are salts are administered parenterally, while the oral dosing is between 3 and 10 times higher than the range previously described.

In addition, the above salts (I) can be used advantageously in experimental polychemotherapy protocols in combination with other antitumor drugs for application generalized clinic in the pathologies described above such as cisplatin, the 5-fluorouracil, vinblastine, cyclophosphamide, Bleomycin, anthracycline and taxol.

Also, the present invention relates to the pharmaceutical compositions containing, as active compound, a therapeutically effective concentration of at least one salt of the formula (I) in combination with excipients and / or diluents adequate.

These pharmaceutical compositions can be prepared in the form of solutions or suspensions, both in media aqueous and non-aqueous, which are particularly effective for their administration in intravenous injections, infusions and subcutaneous or intramuscular injections. These solutions can be prepared before application by dissolving or suspension of lyophilized compounds of the invention in effective solvents.

Similarly, the compositions are indicated solid or semi-solid in the form of implants, gels or ointments for topical, dermal or transdermal administration or in the form of powder, pills, tablets and capsules. In addition, the salts of the invention can be administered in the form of compositions of known controlled release thanks to the state of the art.

The above compositions can be applied easily according to known procedures thanks to state of the art

The following examples, which have no character limited, are described to illustrate the invention:

Experimental part Example 1 Preparation of compound [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2] corresponding to the formula (II) where R 4 = R_ {5} =

 \ break 

methyl

1g of RuCl 3 3 H 2 O (0.0038 was suspended moles) in 30 ml of ethanol and refluxed for three hours until a green solution is obtained Dark. The solution is filtered through a paper to remove the possible traces of undissolved solid; subsequently concentrated until reaching 1/10 of the initial volume, using an evaporator rotary; then 1 ml of concentrated HCl was added 37% aqueous and 2 ml of DMSO. The mixture obtained by this procedure was maintained at a temperature of 80 ° C for approximately 15 minutes, until a color solution is obtained bright orange

After cooling the mixture to room temperature, it is they added 10 ml of acetone; subsequently, the product is separated of the solution in the form of colored crystals oranged Red. The formation of these crystals is accelerated by adding a few drops of ethyl ether. Then, the crystals were collected on a filter, washed successively with cold acetone (20 ml) and with ethyl ether (10 ml) and finally dried under vacuum at room temperature. 1.5 g of the final product with a yield of 72%.

The physicochemical characteristics of [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H] are as follows:

-

Physical state: orange red crystalline solid

-

Gross Formula: C_ {H} {25} Cl_ {O} {4} RuS_ {4}

-

Molecular weight: 556.40

-

Analysis elementary:

Experimental: C = 17.33 H = 4.61 Cl = 25.2 S = 23.2 Theoretical: C = 17.27 H = 4.53 Cl = 25.48 S = 23.04

In addition, the ultraviolet spectra of infrared and structural determination by X-rays coincided with those described by E. Alessio et al. (reference mentioned above).

Example 2 Preparation of [trans-RuCl 4 (DMSO) (Im)] [ImH] corresponding to formula (I) where NR 1 R 2 R 3 is imidazole (Im) and R 4 = R 5 = methyl

1 g (0.0018 mol) of the compound was suspended [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H], prepared according to the procedure described in Example 1, in acetone (20 ml) at temperature environment.

After adding 0.49 g (0.0072 mol) of imidazole, the The mixture was constantly stirred for 4 hours. At this time, the precipitate color gradually turned from orange to red brick. After being collected in a filter and washed successively with acetone (10 ml) and with ethyl ether (10 ml), the product was dried by vacuum at room temperature or in an oven at 60 ° C for some hours. Thus, 0.75 g of final product was obtained with a 92% yield.

The physicochemical properties of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] are the following:

-

Physical state: solid red brick crystal

-

Gross Formula: C_ {H} {15} N_ {4} Cl_ {4} RuS

-

Molecular weight: 458.17

-

Analysis elementary:

Experimental: C = 20.87 H = 3.30 N = 12.2 Theoretical: C = 20.87 H = 3.30 N = 12.23

-

NMR spectrum 1 H of D2S (ppm vs. DSS): -15.2 (very broad, DMSO),

-

3.54 (wide, H2 Im), 7.48 (2, H3 and 4H ImH +), 8.70 (1, H2 ImH +);

-

IR spectrum (selected frequencies, Nujol, cm-1): v_ {NH} 3150 (very wide, medium), v_ {so} 1159 (very strong), v_ {Ru-s} 421 (medium), v_ {Ru-Cl} 342 (strong).

Example 3 Preparation of [trans-RuCl 4 (DMSO) (1-Me-Im)] [1-Me-ImH] corresponding to the formula (I) where NR_R1 {R} {3} is 1-methyl-imidazole and R 4 = R 5 = methyl

1.0 g (0.0018 mol) of the compound was treated [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H], prepared according to the procedure described in Example 1, with 0.59 g (0.0072 mol) of 1-methylimidazole, according to the procedure described in Example 2, and 0.8 g of final product was obtained with a 93% yield.

The physicochemical properties of [trans-RuCl 4 (DMSO) (1-Me-1m)] [1-Me-ImH] are as follows:

-

Physical state: solid red brick crystal

-

Gross Formula: C_ {H} {19} N_ {4} Cl_ {4} RuS

-

Molecular weight: 486.23

-

Analysis elementary:

Experimental: C = 24.8 H = 3.83 N = 11.4 Theoretical: C = 24.7 H = 3.94 N = 11.52

-

NMR spectrum 1 H of D2S (ppm vs. DSS): -15.7 (very broad, DMSO),

-

3.76 (broad, H2 1-Me-Im), -0.95 (broad, Mel, 1-Me-Im) 3.95 (3, Mel, 1-Me-ImH +) 7.45 (2, H3 and 4H 1-Me-ImH +), 8.68 (1, H2 1-Me-ImH +);

-

IR spectrum (selected frequencies, Nujol, cm-1): v_ {NH} 3150 (very wide, medium), v_ {SO} 1093 (very strong), v_ {Ru-S} 424 (medium), v_ {Ru-Cl}, 326 (strong).

Example 4 Preparation of [trans-RuCl 4 (DMSO) (Py)] [PyH] corresponding to formula (I) where NR 1 R 2 R 3 is pyridine and R 4 = R 5 = methyl

1.0 g (0.0018 mol) of the compound was treated [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H], prepared according to the procedure described in Example 1, with 0.57 g (0.0072 mol) of pyridine, according to the procedure described in Example 2. 0.8 were obtained g of the final product with a yield of 94%.

The physicochemical properties of [trans-RuCl_ {4} (DMSO) (Py)] [PyH] are the following:

-

Physical state: dark yellow crystalline solid

-

Gross Formula: C 12 H 17 N 2 Cl 4 ORuS

-

Molecular weight: 480.22

-

Analysis elementary:

Experimental: C = 31.8 H = 3.77 N = 5.43 Theoretical: C = 30.01 H = 3.57 N = 5.83

-

NMR spectrum 1 H of D2S (ppm vs. DSS): -14.5 (very broad, DMSO),

-

2.90 (broad, Py), 8.10, 8.65, 8.82 (PyH +);

-

IR spectrum (selected frequencies, Nujol, cm-1): v_ {NH} 3150 (very wide, medium), v_ {SO} 1074 (very strong), v_ {Ru-s} 432 (medium), v_ {Ru-Cl}, 344 (strong).

Example 5 Preparation of [trans-RuCl 4 (DMSO) (NH3)] [NH4] corresponding to formula (I) where NR 1 R 2 R 3 is NH 3 and R 4 = R 5 = methyl

1.0 g (0.0018 mol) of the compound was suspended [trans- RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H], prepared according to the procedure described in Example 1, in CH 2 Cl 2 (20 ml) a room temperature after emptying by means of a water pump and bubbles of gaseous NH3 were introduced. The system obtained in this way it was kept under stirring for 4 hours, at room temperature, in an ammoniacal atmosphere. During this time, the color of the precipitate gradually turned from orange to red Dark.

The product was collected on a filter and washed successively with cold CH 2 Cl 2 and with ethyl ether (10 ml) and finally dried under vacuum at room temperature.

The physicochemical characteristics of [trans-RuCl 4 (DMSO) (NH 3)] [NH4] are the following:

-

Physical state: solid red brick microcrystalline

-

Gross Formula: C_12 H_ {13} N2 {Cl} {4} ORuS

-

Molecular weight: 456.08

-

Analysis elementary:

Experimental: C = 6.75 H = 3.44 N = 7.63 Theoretical: C = 6.75 H = 3.68 N = 7.87

-

IR spectrum (selected frequencies, Nujol, cm-1): v_ {NH} 3304, 3174 (medium), v_ {SO} 1069 (very strong),

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v_ {Ru-N} 458 (weak), v_ {Ru-S} 431 (medium) v_ {Ru-Cl} 344, 321 (strong).

Example 6 Preparation of [trans-RuCl 4 (DMSO) (Pyr)] [PyrH] corresponding to the formula (I) where NR 1 R 2 R 3 is pyrazine and R 4 = R 5 = methyl

1.0 g (0.0018 mol) of the compound was treated [trans-RuCl 4 (Me 2 SO) 2 H] [(Me 2 SO) 2 H], prepared according to the procedure described in Example 1, with 0.43 g (0.0053 mol) of pyrazine, according to the procedure described in Example 2, and were obtained 0.5 g of final product with a yield of 57%.

The physicochemical properties of [trans-RuCl 4 (DMSO) (Pyr)] [PyrH] are the following:

-

Physical state: dark yellow crystalline solid

-

Gross Formula: C 10 H 15 N 4 Cl 4 ORuS

-

Molecular weight: 482.20

-

Analysis elementary:

Experimental: C = 24.7 H = 3.07 N = 11.1 Theoretical: C = 24.9 H = 3.13 N = 11.6

-

Spectrum of 1 H NMR of D2S (ppm vs. DSS): -13.8 (broad, DMSO),

-

7.5 (very wide, H2, 6 Pyr), -2.1 (broad, H3.5 Pyr) 8.69 (Pyr);

-

IR spectrum (selected frequencies, Nujol, cm-1): v_ {Pirazina} 1605 (medium), v_ {SO} 1076 (very strong), v_ {Pirazina} 807 (strong), v_ {Pirazina \; protonated} 773 (strong) v_ {Ru-S} 435 (medium), v_ {Ru-Cl} 351,326 (strong).

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UV / VIS in water (nm, ε (mol-1 cm-1): 400 (4200), 469 (490).

Biological activity i) Tests of the in vivo activity of salts (I), according to the present invention, in mice affected by carcinoma Lewis lung.

Example (i) -A

Material and Procedures

Three groups, each consisting of 10 mice BD2F1 female of 2 ± 1 g obtained from the Charles River (Calco, Como, Italy), were inoculated by intramuscular injection with 10 6 Lewis lung carcinoma cells suspended in 0.05 ml of phosphate buffered magnesium and calcium free solution Dulbecco (PBS) using a sterile insulin syringe.

The tumor line was obtained from the Tumor Repository Bank, NCl, NIH of Bethesda (USA) and remained in nitrogen liquid.

Five days after said inoculation, the weight average of the tumors was 0.4 ± 0.01 g. From the fifth and eleventh day, the three groups of mice were treated intraperitoneally as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] Na 2DMSO,  as mentioned in the international patent application WO 90/13553, cited above;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

After 12 days from the inoculation, the primary tumor was removed by surgery.

After 21 days from the inoculation, mice were sacrificed by dislocation cervical and lung metastases were counted. The lungs are they separated immediately after the death of the mice and they divided into unique lobes, which were subsequently examined by using a stereoscopic microscope of great magnification, equipped with a grid in the eyepiece that allowed the detection of orthogonal axes a and b (in which a \ leqb).

Pulmonary metastases were classified subsequently according to the dimensions and then it calculated the weight of each animal's metastases as the sum of the weight of each metastasis, considered individually as a solid, by the formula (? / 6) a2 xb. The data Experimental obtained were finally processed with the test Student-Newmann-Keuls statistic.

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media \pm error típico (E.R.) de los valores aislados obtenidos para cada grupo.The results obtained are shown in Table 1 below, in which the number of metastases and weight in groups (2) and (3) of the mice treated with the Control Group (1) is compared. The data is shown as the

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mean ± standard error (ER) of the isolated values obtained for each group.

TABLE 1 Antitumor and antimetastatic activity of [trans-RuCl 4 (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) versus [trans-RuCl 4 (DMSO) (Im)] [ ImH] Na 2DMSO (44 mg / kg / day) in mice affected by

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Lewis lung carcinoma

Compound No. of metastases per animal Weight of animal metastases Control 28.9 ± 5.2 16.3 ± 5.2 [trans-RuCl 4 (DMSO) (Im)] 11.9 ± 2.2 * 2.8 ± 0.8 * [ImH] 2DMSO [trans-RuCl 4 (DMSO) (Im)] 8.8 ± 1.3 * 2.2 ± 0.3 * [ImH] * p <0.05 front to the control group; t test for data grouped

The data shown above underline the fact that treatment with [trans-RuCl_ {4} (DMSO) (Im)] [ImH], of according to the present invention, causes a reduction of both the weight as of the number of metastases higher than that obtained with the known reference compound thanks to the state of the technique.

The percentage reduction (%) in the number of metastasis and its weight is shown below in the Table two.

TABLE 2 Reduction percentage (%) in the number of metastasis and weight in mice affected by carcinoma Lewis lung and treated with the compounds according to the present invention

Compound (%) Reduction in the number of metastasis (%) Reduction in the number of metastases [trans-RuCl 4 (DMSO) (Im)] 58.3 83.0 [ImH] Na 2DMSO [trans-RuCl 4 (DMSO) (Im)] 69.1 86.4 [ImH]

The advantages of the compound of the invention against the reference compound, as known in the state of the art, are indicated more clearly with the data shown below.

After the previous treatments, the metastases isolated from the groups of mice (1) and (3) were counted and they divided into three groups depending on the dimension of their diameter (d): metastases with a diameter d <1 mm (small), metastases with a diameter ranging from 1 mm to 2 mm (medium) and, finally, metastases with a diameter d> 2 mm (large). The Results obtained are shown below in Table 3.

TABLE 3 Distribution according to the dimensions of the diameter (d) of lung metastases in mice affected by Lewis lung carcinoma, treated with the compounds of the present invention

Compound Metastasis of d <1 mm 1mm metastasis <d <2mm Metastasis d> 2mm Control 68.4% 29.8% 1.8% [trans-RuCl 4 (DMSO) (Im)] 83.0% 17.0% 0% [ImH]

These data underline the fact that the nodules metastatic of medium and large dimensions of the group of animals treated are scarcely present (nodules of dimensions means) or absent (large nodules) in relation to Control animals

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Example (i) -B

Material and procedures

Three groups, each consisting of 10 mice BD2F1 female of 21 ± 1 g obtained from the Charles River (Calco, Como, Italy), were inoculated by intramuscular injection with 10 6 Lewis lung carcinoma cells suspended in 0.05 ml of phosphate buffered magnesium and calcium free solution Dulbecco (PBS), using a sterile insulin syringe.

The tumor line was obtained from the Tumor Repository Bank, NCl, NIH of Bethesda (USA) and remained in nitrogen liquid.

Five days after said inoculation, the weight average of the tumors was 0.4 ± 0.01 g. From twelfth and seventeenth day, the three groups of mice were treated intraperitoneally as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 2 mg of cisplatin;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

After 11 days from the inoculation, the primary tumor was removed by surgery.

After 25 days from the inoculation, mice were sacrificed by dislocation cervical and lung metastases were counted. The lungs are they separated immediately after the death of the mice and they divided into unique lobes, which were subsequently examined by using a stereoscopic microscope of great magnification, equipped with a grid in the eyepiece that allowed the detection of orthogonal axes a and b (in which a \ leqb).

Pulmonary metastases were classified subsequently according to the dimensions and then it calculated the weight of each animal's metastases as the sum of the weight of each metastasis, considered individually as a solid, by the formula (? / 6) a2 xb. The data Experimental obtained were finally processed with the test statistical Student-Newmann-Keuls.

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media ± error típico (E.R.) de los valores aislados obtenidos para cada grupo.The results obtained are shown in Table 4 below, in which the number of metastases and weight in groups (2) and (3) of the mice treated with the Control Group (1) is compared. The data is shown as the

 \ break 

mean ± standard error (ER) of the isolated values obtained for each group.

TABLE 4 Antimetastatic activity of [trans-RuCl 4 (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) against cisplatin

 \ break 

(2 mg / kg / day) in mice affected by Lewis lung carcinoma

Compound No. of metastases per animal Weight of lung metastases (mg) Control 14.8 ± 0.6 985.0 ± 41.0 [trans-RuCl 4 (DMSO) (Im)] 4.3 ± 0.5 * 73.3 ± 20.0 * [ImH] Cisplatin 13.5 ± 0.6 * 865.1 ± 72.8 * * p <0.05 front to control against cisplatin; t test for data grouped

The data shown above underline the fact that treatment with [trans-RuCl_ {4} (DMSO) (Im)] [ImH] of according to the present invention causes a reduction in both the weight as in the number of metastases higher than that obtained with the reference compound.

Example (ii) -A

Material and procedures

Three groups of 7 female CBA mice of 23 ± 2 g, obtained from the growth of a colony according to the procedures for related animals. The colony obtained originally from the Chester Beatthy Institute in London it subsequently maintained by serial crossings between relatives (brother and sister) in a ratio that varied between 1: 1 and 1: 4. The crosses coincided with the maximum sexual maturity of the Animals born in the fifth week. Animals with a weight less than normal or with obvious organic abnormalities were discarded The three groups were inoculated by injection intramuscular with 10 6 Lewis lung carcinoma cells suspended in 0.05 ml of magnesium and calcium free solution buffer with Dulbecco phosphates (PBS), using a syringe sterile insulin

The tumor line was obtained from Rudjer Boskovic Institute and remained in liquid nitrogen.

After 13 days from the Inoculation, the average weight of the tumors was 1.2 ± 0.2 g. From the thirteenth day to the eighteenth day, the three groups of mice were treated intraperitoneally as follows way:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 44 mg of [trans-RuCl 4 (DMSO) (Im)] Na 2DMSO, according to It is mentioned in international patent application WO 90/13553, cited above;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

After 19 days from the inoculation, the primary tumor was removed by surgery.

After 27 days from the inoculation, mice were sacrificed by dislocation cervical and lung metastases were counted, as described in the experiment (i).

The experimental data obtained were processed finally with the statistical essay Student-Newmann-Keuls.

In order to evaluate the activity of salts of the invention, the primary tumor of the the mice of the three previous groups and subsequently dissected One tumor was randomly chosen from each group, from which 10 sections / tumor were obtained with the intention of assessing the mass total neoplastic Tumor cuts were fixed in formalin at 10%, were embedded in paraffin and stained with dye Cajal-Gallego to observe the cells epithelial, connective matrix and the presence of erythrocytes. The infiltration was evaluated by observing the degranulation of the polymorphonuclear leukocytes (PMN), as well as the presence of Apoptotic bodies.

Results

The results obtained are shown in the Table 5, which compares the number of metastases and their weight obtained in groups (2) and (3) with the Control Group (1). The data is show as the mean ± typical error (E.T.) of the values isolates obtained for each group.

TABLE 5 Antitumor and antimetastatic activity of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) versus [trans-RuCl_ {4} (DMSO) (Im)] [ImH] Na 2DMSO (44 mg / kg / day) in mice affected by breast carcinoma (MCa)

Compound No. of metastases per animal Weight of animal metastases Control 39.2 ± 8.1 31.4 ± 10.2 [trans-RuCl 4 (DMSO) (Im)] 35.2 ± 5.5 8.0 ± 2.5 * [ImH] 2DMSO [trans-RuCl 4 (DMSO) (Im)] 6.8 ± 2.2 * 0.5 ± 0.2 ** [ImH] * p <0.05 front to the control group; ** p <0.05 versus the control group and p <0.05 in front of the group (2) treated with [trans-RuCl_ {4} (DMSO) (Im)] [ImH] 2DMSO Na; test of t for grouped data.

The percentage reduction in the number of metastasis and in its weight, achieved with the compound of the invention and with the known sodium salt thanks to the state of the technique versus control, shown below in the Table 6.

TABLE 6 Reduction percentage (%) in the number of metastasis and in its weight achieved with the compounds according with the present invention

Compound (%) Reduction in the number of metastasis (%) Reduction in the weight of metastasis [trans-RuCl 4 (DMSO) (Im)] 10.2 74.5 [ImH] Na 2DMSO [trans-RuCl 4 (DMSO) (Im)] 82.6 98.4 [ImH]

The results related to the effect of salts of the invention are shown below in Table 7.

TABLE 7 Effects of the compounds according to the present invention on the primary tumor

Control [trans-RuCl 4 (DMSO) (Im)] [trans-RuCl 4 (DMSO) [ImH] 2DMSO (Im)] [ImH] (44 mg / kg / day) (35 mg / kg / day) Zones of hemorrhage Limited Discreet Numerous and very wide Zones of necrosis Limited discreet Very wide Condition of infiltration High, PMN intact High, with PMN partially Clear degranulation of the degranulates and Some PMN; full from bodies apoptotic apoptotic bodies Connective capsule Normal, with beams Limited in thickness and Wide, thick and infiltrating muscle do not hard to distinguish the tissue tumor always intact of the muscle layer

According to the data shown above, it is possible to observe that the compound [trans-RuCl_ {4} (DMSO) (Im)] [ImH], versus to the reference compound [trans-RuCl_ {4} (DMSO) (Im)] [ImH] Na 2DMSO, 44 mg / kg / day, causes deep histological changes. In in particular, the tumor treated with salt according to the present invention has a greater area of hemorrhage and necrosis combined with neutrophil leukocytes, a clear degradation of the polymorphonuclear leukocytes (PMN), a high presence of apoptotic bodies and an increase in the thickness of the connective capsule surrounding the tumor

\hbox{B2 y
C2).}

These results are even more evident in the figures in which the outer connective capsule is designated with the letter C (Figure Al, B1 and C1), while the area of necrosis is designated with the letter N (Figure A2,

 \ hbox {B2 and
C2).} 

Without the intention of restricting the mechanism of action with which salts, according to the present invention, exert their pharmacological effects, the previous observations they contribute to explain, at least partially, the effect antimetastatic of the above salts as the result of the prevention of tumor cell reproduction from primary tumor

Example (ii) -B

Material and procedures

Two groups of 12 mice and one group (control) of 14 female CBA mice of 22 g, obtained from growth from a colony according to animal procedures related, inoculated with 10 6 carcinoma cells of breast (MCa), as shown in the experiment (ii) -A.

From the ninth to the fourteenth day later of inoculation, the three groups of mice were treated intraperitoneally as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 27 mg of [trans-RuCl 4 (DMSO) (NH 3)] [NH4] (Example 5).

After 27 days from the inoculation, mice were sacrificed by dislocation cervical and lung metastases were counted, as described in the experiment (i).

The experimental data obtained were processed finally with the statistical essay Student-Newmann-Keuls.

In order to evaluate the activity of salts of the invention on the primary tumor, the average weight was calculated of the primary tumor at the beginning and end of treatment previous.

Results

The results obtained are shown in the Table 8, which compares the number of metastases and their weight obtained in groups (2) and (3) with the Control Group (1). The data is show as the mean ± typical error (E.T.) of the values isolates obtained for each group.

TABLE 8 Antitumor and antimetastatic activity of [trans-RuCl 4 (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) and of [trans-RuCl 4 (DMSO) (NH 3) )] [NH4] described in Example 5 (27 mg / kg / day) in mice affected by

 \ break 

breast carcinoma (MCa)

Compound No. of metastases per animal Weight of animal metastases Control 40.4 ± 6.0 115.0 ± 22.1 [trans-RuCl 4 (DMSO) (Im)] 20.7 ± 4.0 * 23.2 ± 7.2 ** [ImH] [trans-RuCl 4 (DMSO) 15.6 ± 3.2 * 10.7 ± 2.2 ** (NH3)] [NH4] * p <0.05 and ** p <0.01 versus control group; t test for data grouped

The percentage reduction (%) in the number of metastasis and in its weight, achieved with the compound of the invention and with the known sodium salt thanks to the state of the technique versus control, shown below in the Table 9.

TABLE 9 Reduction percentage (%) in the number of metastasis and its weight, obtained with the compounds according to the present invention

Compound (%) Reduction in the number of metastasis (%) Reduction in the weight of metastasis [trans-RuCl 4 (DMSO) (Im)] 48.8 79.8 [ImH] [trans-RuCl 4 (DMSO) 61.4 90.7 (NH3)] [NH4]

The results related to the effect of salts of the invention on the growth of the primary tumor are shown in Table 10, in which the tumor weight is indicated primary at the beginning and at the end of the treatment with the salts of the invention, as well as the corresponding percentage of increase.

TABLE 10 Effects exerted by the compounds according to the present invention on the growth of the primary tumor in mice affected by breast carcinoma (MCa)

Tumor weight primary (mg) Compound Treatment start End of the treatment Increase (%) Control 157 ± 16 723 ± pm98 +461 [trans-RuCl 4 (DMSO) (Im)] [ImH] 152 ± 19 282 ± 61 ** +185 [trans-RuCl 4 (DMSO) (NH3)] [NH4] 191 ± 23 490 ± 91 * +256 * p <0.05 and ** p <0.01 versus the control group; t test for data grouped

Based on the data shown above, the reduction of primary tumor growth in treated groups with the salts according to the present invention, standardized for controls, it is 59.9% for the [trans- RuCl_ {4} (DMSO) (Im)] [ImH], while for [trans-RuCl 4 (DMSO) (NH 3)] [NH4] is 44.5%.

Example (ii) -C

Material and procedures

Two groups of 7 mice and a group of 9 (controls) 22 g female CBA mice, obtained from growth of a colony according to the procedures for related animals, were inoculated with 10 6 cells of breast carcinoma (MCa) by intramuscular injection, as sample in experiment (ii) -A.

From the ninth day to the fourteenth day, the three groups of mice were treated daily by route intraperitoneal as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 44 mg of [trans-RuCl 4 (DMSO) (Im)] Na 2DMSO, according to It is mentioned in international patent application WO 90/13553, cited above;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 27 mg of [trans-RuCl 4 (DMSO) (NH 3)] [NH4] (Example 5).

In order to evaluate the activity of salts of the invention on the primary tumor, the average weight was calculated of the primary tumor at the beginning and end of treatment with salts of the invention, as well as the percentage variation of the increase.

The results obtained are shown at continued in Table 11, which indicates the weight of the primary tumor at the beginning and at the end of the treatment, as well as the variation of increase (%).

TABLE 11 Effect exerted by the compounds according to the present invention on the growth of the primary tumor of mice affected by breast carcinoma (MCa)

Tumor weight primary (mg) Compound Treatment start End of the treatment Increase (%) Control 355 ± 31 813 ± 1010 +229 [trans-RuCl 4 (DMSO) (NH3)] 367 ± 46 681 ± 62 +186 Na 2DMSO [trans-RuCl 4 (DMSO) (NH 3)] [NH4] 410 ± 17 633 ± 38 * +154 * p <0.05 front to the control group; t test for data grouped

Based on the data shown above, evidence that primary tumor growth is reduced by those groups treated with the compound salts of Ru (III), normalized by controls, at 18.7% for [trans-RuCl 4 (DMSO) (NH 3)] Na 2DMSO, while for the [trans-RuCl 4 (DMSO) (NH 3)} [NH4] is reduced by 32.8%. These data show that the [trans- RuCl 4 (DMSO) (NH 3)] [NH 4] of the The present invention may exert anti-tumor effects on the primary tumor growth, indicating a reduction well above that achieved by known compounds thanks to the state of the art.

Example (ii) -D

Material and procedures

Three groups of 7 female CBA mice of 23 ± 2 g were obtained from the growth of a colony according with procedures for related animals. The colony originally obtained from the Chester Beatthy Institute in London it was subsequently maintained by serial crossings between relatives (brother and sister) in a ratio that varied between 1: 1 and 1: 4. The crosses coincided with the maximum sexual maturity of the Animals born in the fifth week. Animals with a weight less than normal or with obvious organic abnormalities were discarded The three groups were inoculated by injection intramuscular with 10 6 Lewis lung carcinoma cells suspended in 0.05 ml of magnesium and calcium free solution buffer with Dulbecco phosphates (PBS), using a syringe sterile insulin

The tumor line was obtained from Rudjer Boskovic Institute and remained in liquid nitrogen.

Thirteen days after inoculation, it was removed by surgery the primary tumor.

From the fourteenth day to the day Nineteenth, the three groups of mice were treated intraperitoneally as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 2 mg of cisplatin;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

TABLE 12 Comparison of the antimetastatic effect of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) against cisplatin (2 mg / kg / day) in mice affected by breast carcinoma (MCa)

Compound Effects on metastasis pulmonary Days of survival (mean \ typical pmerror) Control 17.4 ± 1.2 [trans-RuCl_ {4} (DMSO) (Im)] [ImH] 24.4 ± 2.2 * Cisplatin 24.3 ± 1.7 * * p <0.05 versus controls

Example (ii) -E

In vivo experiment of salt activity (I), according with the present invention, in mice affected by the breast adenocarcinoma (TS / A) Material and procedures

Three groups each consisting of 10 mice BALBc female of 21 ± 1 g, obtained from Harlan Nossan (Italy), is inoculated by intramuscular injection with 10 5 cells of breast adenocarcinoma (TS / A) suspended in 0.05 ml of solution free of calcium and magnesium buffered with Dulbecco phosphates (PBS), using a sterile insulin syringe.

The tumor line was obtained from the Institute immunogenetics (Univ. Torino, IT) and remained in nitrogen liquid.

From the thirteenth day to the day Eighteenth, the three groups of mice were treated intraperitoneally as follows:

Group 1 - Control: 10 ml / kg body weight / day of a non-sterile and sterile physiological solution;

Group 2 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 2 mg of cisplatin;

Group 3 - 10 ml / kg body weight / day of one physiogenic and sterile physiological solution containing 35 mg of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] (Example two);

After 19 days from the inoculation, the primary tumor was removed by surgery.

After 33 days from the inoculation, mice were sacrificed by dislocation cervical and lung metastases were counted. The lungs are they separated immediately after the death of the mice and they divided into unique lobes, which were subsequently examined by using a stereoscopic microscope of great magnification, equipped with a grid in the eyepiece that allowed the detection of orthogonal axes a and b (in which a \ leqb).

Pulmonary metastases were classified later depending on the dimensions and then it calculated the weight of each animal's metastases as the sum of the weight of each metastasis, considered individually as a solid, by the formula (? / 6) a2 xb. The data Experimental obtained were finally processed with the test statistical Student-Newmann-Keuls.

The results obtained are shown at then in Table 13, in which the number of metastasis and weight in groups (2) and (3) of the mice treated with the Control Group (1). The data is shown as the mean ± standard error (E.T.) of the isolated values obtained For each group.

TABLE 13 Antimetastatic activity of [trans-RuCl 4 (DMSO) (Im)] [ImH] of Example 2 (35 mg / kg / day) against cisplatin

 \ break 

(2 mg / kg / day) in mice affected by breast adenocarcinoma TS / A

Compound No. of metastases per animal Weight of lung metastases (mg) Control 7.3 ± 3.0 10.4 ± 3.1 [trans-RuCl 4 (DMSO) (Im)] 1.7 ± 0.5 * 0.4 ± 0.1 * [ImH] Cisplatin 3.70 ± 1.0 * 1.6 ± 0.6 * * p <0.05 front to control; t test for pooled data

The treatments chosen for the comparison of the effects of [trans-RuCl_ {4} (DMSO) (Im)] [ImH] and the Cisplatin on metastatic solid tumors are comparable since they represent the maximum tolerated doses with the treatment adopted (1 injection / day / for 6 days in a row).

The comparison of the effects of the compound of the invention and cisplatin on the reduction of accumulated weight during treatment shows that the ruthenium compound (percentage (%) of accumulated weight reduction versus control = -R1 (Lewis lung carcinoma), -6 (MCa breast carcinoma) is always less toxic than cisplatin (percentage (%) of weight accumulated against the control = -11). Spleen weight comparison of treated animals (percentage (%) of weight reduction of spleen versus control = -11) also shows that treatment with the compound of the invention is much more tolerant than the Cisplatin treatment on animals (percentage (%) of reduction of spleen weight versus control = -52).

In tumor metastasis, the compound of the invention is effective, in the case of breast carcinoma (MCa) and even more effective, in the case of Lewis lung carcinoma and TS / A adenocarcinoma, than cisplatin. It also reduces the number of lung metastases even when administered during the early stages of tumor growth, that is, before  of surgical ablation (TS / A adenocarcinoma) or when administered in mice with lung metastases during phases of advanced growth, that is, after surgical ablation of the primary tumor (Lewis lung carcinoma). The reduction of pulmonary metastases entails a significant prolongation of postoperative life expectancy of treated mice (MCa breast carcinoma).

The experimental in vivo models shown previously refer to the treatment, with the compounds of the present invention of two solid extended tumors in rodents in a phase of advanced growth. A reduction is observed important and statistically significant both in growth of the primary tumor as in the formation of pulmonary metastases. This reduction, as can be seen in model (i) of Lewis lung carcinoma and in model (ii) of carcinoma of MCa breast is attributed to a pronounced growth inhibition of lung metastases. In fact, in relation to control animals, the presence of large nodules / stockings in the treated animal groups is notably inferior and even non-existent in terms of nodules big.

In particular, from the set of comparisons of data that refer to effective reduction In metastases, the compounds of the present invention are Unusually more active than known normal compounds thanks to the state of the art, both in the treatment of Lewis lung carcinoma as, more clearly and statistically significant, in the treatment of carcinoma of breast (MCa).

Claims (24)

1. A salt of an anionic compound of Ru (III) with an ammonium cation of formula (I): one where R 1, R 2 and R 3, the same or different from each other, they are selected from a group formed by H, C 1 -C 6 alkyl, linear or branched, saturated or unsaturated, C 3 -C 7 cycloalkyl, phenyl and aryl; or NR 1 R 2 R 3 is a heterocycle of 5-7 members formed by nitrogen, saturated or unsaturated, optionally containing one or more atoms of O, S and / or N; said nitrogen atom being optionally substituted by a residue C 1 -C 4 alkyl, aryl or benzyl; said nitrogen containing heterocycle being condensed optionally with a benzyl group and / or replaced by a group C 1 -C 4 alkyl, alkoxy C 1 -C 4, aryl or benzyl; where R_ {4} and R_ {5}, same or different each other, they are selected from the group consisting of an atom of H, a C1-C6 alkyl, a cycloalkyl C3-C7, a phenyl and an aryl or R4 and R6 form, together with the atom of S, a heterocycle of 4-7 members. 2. The salt according to claim 1, characterized in that R1, R2 and R3 are H or ethyl. 3. The salt according to claim 1, characterized in that NR1R2R3 is a nitrogenous 5-membered heterocycle selected from the group consisting of imidiazole, N-methylimidazole, pyrazole and oxazole 4. The salt according to claim 1, characterized in that NR1R2R3 is a 6-membered heterocycle selected from the group consisting of pyridine, pyrazine, 3,5-lutidine and 4 -methylpyridine. 5. The salt according to claim 1, characterized in that NR1R2R3 is a nitrogen heterocycle that is condensed with a benzyl group selected from indazole, isoqinoline, benzimidazole and 1.5 , 6-trimethyl-benzimidazole. 6. The salt according to claims 1-5, characterized in that R4 -SO-R5 is selected from the group consisting of dimethylsulfoxide, diethylsulfoxide, tetramethylene sulfoxide. 7. The salt according to claims 1, 3 and 6, characterized in that NR1R2R3 is imidazole and

 \ break 

R 4 -SO-R 5 is dimethylsulfoxide. 8. The salt according to claims 1, 2 and 6, characterized in that NR 1 R 2 R 3 is NH 3 and R 4 -SO-R 5 is dimethylsulfoxide. 9. Procedure for the preparation of a salt of formula (I) according to claims 1-8, which It comprises the following reaction steps:

one)

RuCl_ {3} reacts with R4 -SO-R5 in presence of HCl to produce a compound of formula (II):

two where R_ {4} and R_ {5} have the meaning before indicated.

two)

The compound (II), obtained from step (1), reacts at room temperature with a nitrogenous compound, NR 1 R 2 R 3, in one or more organic solvents, to produce the salt of formula (I).

10. The process according to claim 9, characterized in that R4 and R5 are methyl or ethyl. 11. The process according to claim 9, characterized in that, in step (1), RuCl 3 is dissolved in an organic solvent and the solution obtained by this process reacts with R 4 -SO- R 5 and concentrated HCl at a temperature ranging between 60 ° C and 90 ° C. 12. The process according to claim 11, characterized in that the RuCl 3 is dissolved in ethanol or methanol and the solution obtained by this procedure reacts with R 4 -SO-R 5 and concentrated HCl at an approximate temperature of 80 ° C. 13. The process according to claim 9, characterized in that, in step (2), said compound (II) and said nitrogen compound, NR1R2R3, react in molar ratios ranging from 1: 2 to 1: 6. 14. The process according to claim 9, characterized in that, in step (2), said organic solvents are acetone and dichloromethane. 15. The use of at least one of the salts of formula (I) according to any one of the claims 1-8, in the manufacture of a medicine that has antimetastatic and antineoplastic activity. 16. The use according to claim 15 in the that said medication is supplied for the treatment of tumors metastatic solids. 17. The use according to claim 16, characterized in that said solid tumor is selected from the group consisting of carcinomas of the gastroinensayoinal tract, breast carcinomas, lung tumors, metastatic carcinomas, pulmonary metastases of metastatic tumors. 18. The use according to claim 15 in the that said medication is effective for dose administration between 0.1 and 300 mg / kg / day of said salts. 19. The use according to claim 15, characterized in that said medicament is effective for parenteral, oral, topical or transdermal administration. 20. A pharmaceutical composition containing, as an active compound, an effective concentration therapeutically of at least one of the salts of formula (I), as defined in claims 1-8, in combination with suitable excipients and diluents. 21. The pharmaceutical composition according to claim 20, characterized in that it is in the form of a solution or suspension. 22. The pharmaceutical composition according to claim 20, characterized in that it is in the form of a gel, ointment, powder, pill, tablet, capsule or implant. 23. The pharmaceutical composition according to claim 20, characterized in that said salts of formula (I) are combined with one or more antitumor drugs. 24. The pharmaceutical composition according to claim 23, characterized in that said antitumor drugs are selected from the group consisting of cisplatin, 5-fluorouracil, vinblastine, cyclophosphamide, bleomycin, anthracycline and taxol.